Human adipose-derived stem cells enriched with VEGF-modified mRNA promote angiogenesis and long-term graft survival in a fat graft transplantation model.
Human adipose-derived stem cells enriched with VEGF-modified mRNA promote angiogenesis and long-term graft survival in a fat graft transplantation model.
复制标题
富含 VEGF 修饰 mRNA 的人类脂肪干细胞可促进脂肪移植模型中的血管生成和长期移植物存活。
DOI：
10.1186/s13287-020-02008-8
复制
发表时间：
2020-11-19
影响因子：
7.5
通讯作者：
Fu Y
中科院分区：
文献类型：
--
作者：
Yu F;Witman N;Yan D;Zhang S;Zhou M;Yan Y;Yao Q;Ding F;Yan B;Wang H;Fu W;Lu Y;Fu Y
脂肪移植作为许多软组织缺陷的标准治疗方法，仍然是不可预测的，并且依赖于技术。人类脂肪干细胞(HADSCs)是细胞辅助治疗提高移植物存活率的有希望的候选细胞。由于自由生活脂肪需要营养和呼吸源才能蓬勃发展，血管形成不足和不稳定仍然阻碍了hADSC辅助治疗。近年来，细胞疗法联合编码血管内皮生长因子(VEGF)的修饰信使核糖核酸(ModRNA)已被应用于缺血性疾病的治疗。在此，我们假设，在脂肪移植模型中，血管内皮生长因子modRNA(ModVEGF)工程的hADSCs可以显著提高脂肪存活率。从脂肪抽吸法获得hADSC，并将modRNAs导入hADSCs。首次在体外评价了modRNAs在hADSCs中的转染率和表达动力学。接下来，我们应用体内Matrigel Plug试验来评估植入后1 周的modVEGFhADSCs的活性和血管生成潜能。最后，在小鼠模型中，将modVEGF工程的hADSCs与人类脂肪共移植，以分析长期随访中脂肪移植物的存活率、再血管化、增殖、纤维化、凋亡和坏死。在我们的体外和体内模型中，modVEGF在hADSCs中的转移是高度耐受的，因为modVEGF工程的hADSCs促进了VEGF的突发性蛋白的产生。在体外和体内实验中，与未经处理的hADSCs相比，modVEGF工程的hADSCs诱导了更多的细胞增殖和促血管生成。在脂肪移植模型中，我们提供了证据表明，modVEGF工程的hADSCs促进了保存脂肪细胞的最佳效力，特别是在移植后的长期阶段。对体内移植后15、30和90 天收获的脂肪移植物的详细组织学分析表明，从modVEGF工程的hADSC中释放血管内皮生长因子蛋白显著促进了新生血管生成、血管成熟度和细胞增殖。与对照组相比，modVEGF工程的hADSCs还显著减轻了移植物的纤维化、凋亡和坏死。此外，modVEGF工程的hADSCs促进了移植物的存活和细胞分化能力，这也诱导了移植后血管形成和存活脂肪细胞数量的增加。目前的研究表明，利用modVEGF工程的hADSCs作为临床治疗的高级替代方案，涉及软组织重建和年轻化。
Fat grafting, as a standard treatment for numerous soft tissue defects, remains unpredictable and technique-dependent. Human adipose-derived stem cells (hADSCs) are promising candidates for cell-assisted therapy to improve graft survival. As free-living fat requires nutritional and respiratory sources to thrive, insufficient and unstable vascularization still impedes hADSC-assisted therapy. Recently, cytotherapy combined with modified mRNA (modRNA) encoding vascular endothelial growth factor (VEGF) has been applied for the treatment of ischemia-related diseases. Herein, we hypothesized that VEGF modRNA (modVEGF)-engineered hADSCs could robustly enhance fat survival in a fat graft transplantation model. hADSCs were acquired from lipoaspiration and transfected with modRNAs. Transfection efficiency and expression kinetics of modRNAs in hADSCs were first evaluated in vitro. Next, we applied an in vivo Matrigel plug assay to assess the viability and angiogenic potential of modVEGF-engineered hADSCs at 1 week post-implantation. Finally, modVEGF-engineered hADSCs were co-transplanted with human fat in a murine model to analyze the survival rate, re-vascularization, proliferation, fibrosis, apoptosis, and necrosis of fat grafts over long-term follow-up. Transfections of modVEGF in hADSCs were highly tolerable as the modVEGF-engineered hADSCs facilitated burst-like protein production of VEGF in both our in vitro and in vivo models. modVEGF-engineered hADSCs induced increased levels of cellular proliferation and proangiogenesis when compared to untreated hADSCs in both ex vivo and in vivo assays. In a fat graft transplantation model, we provided evidence that modVEGF-engineered hADSCs promote the optimal potency to preserve adipocytes, especially in the long-term post-transplantation phase. Detailed histological analysis of fat grafts harvested at 15, 30, and 90 days following in vivo grafting suggested the release of VEGF protein from modVEGF-engineered hADSCs significantly improved neo-angiogenesis, vascular maturity, and cell proliferation. The modVEGF-engineered hADSCs also significantly mitigated the presence of fibrosis, apoptosis, and necrosis of grafts when compared to the control groups. Moreover, modVEGF-engineered hADSCs promoted graft survival and cell differentiation abilities, which also induced an increase in vessel formation and the number of surviving adipocytes after transplantation. This current study demonstrates the employment of modVEGF-engineered hADSCs as an advanced alternative to the clinical treatment involving soft-tissue reconstruction and rejuvenation.
登录
查看更多内容
影响因子：
168.9
作者：
Kolle, Stig-Frederik Trojahn;Fischer-Nielsen, Anne;Drzewiecki, Krzysztof Tadeusz
通讯作者：
Drzewiecki, Krzysztof Tadeusz
影响因子：
2.9
作者：
Harris, William M.;Plastini, Michael;Zhang, Ping
通讯作者：
Zhang, Ping
影响因子：
5.3
作者：
Li J;Chen S;Zhao Z;Luo Y;Hou Y;Li H;He L;Zhou L;Wu W
通讯作者：
Wu W
影响因子：
3.6
作者：
Fu, Su;Luan, Jie;Gao, Yunzhou
通讯作者：
Gao, Yunzhou
影响因子：
10.8
作者：
Diaz-Herraez, Paula;Saludas, Laura;Jose Blanco-Prieto, Maria
通讯作者：
Jose Blanco-Prieto, Maria